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Fig. 4. Neurosphere cultures from Gli2 null brains. (A-D) Phase contrast
images of representative cortical nsps cultured from wild-type (A,B) and
Gli2-/- (C,D) animals at E18.5. (E) RT-PCR analyses of
cortical nsps. Note the loss of Gli1 expression, the shift in the
Gli2 mutant allele band (arrows), the reduced Gli3
expression, and the induction of Ihh and to a lesser extent
Dhh in Gli2-/- cells. Hprt is shown as a
control. (F-I) Expression of nestin in precursors of
Gli2-/- nsps (F), of TuJ1 in neurons (G), of GFAP in
astrocytes (H) and of O4 in oligodendrocytes (I) after differentiation. Nuclei
were counterstained with DAPI. (J-M) Quantification of nsp size at E15.5 (J)
and E18.5 (K). An average of 20 nsps from two independent experiments is shown
(E15.5, P<0.05; E18.5, P<0.001). Given their rarity at
E18.5, single Gli2-/- nsps were measured. (L,M)
Quantification of nsps obtained in cloning assays. Three independent
experiments are shown for E15.5 (P<0.001) and one for E18.5. Scale
bar: 75 µm for A-D; 10 µm for F-I.
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